INVESTIGATION  OF  THE  ALKALOID  CONTENT  OF  SOME 

SOUTH  AMERICAN  BARKS 


By 


MAXIMO  ELADIO  MORALES 


THESIS 


FOR  THE 

DEGREE  OF  BACHELOR  OF  SCIENCE 
IN  CHEMISTRY 


COLLEGE  OF  LIBERAL  ARTS  AND  SCIENCES 
UNIVERSITY  OF  ILLINOIS 


1921 


**x 


19 1\ 


UNIVERSITY  OF  ILLINOIS 


U 

T 


X) 


THIS  IS  TO  CERTIFY  THAT  THE  THESIS  PREPARED  UNDER  MY  SUPERVISION  BY 



ENTITLED HI  YE  SX  IGAXI  M.  _ JliU  H^HE.jUxKAIlQ  IH  _ U OUTER  T_  _QZ  _ CJME. 

SOUlH_j^M£lBXCAlL_BiiBjcCS_._ 

IS  APPROVED  BY  ME  AS  FULFILLING  THIS  PART  OF  THE  REQUIREMENTS  FOR  THE 

degree  of  B.a5JieJlox_XLf_^cJLe2iQ£._in_I2hemia.tiy 


Instructor  in  Charge 


HEAD  OF  DEPARTMENT  OF 


47G6G4 


INTRODUCTION 

The  research  work  that  I have  done  embraces  a period  of 
three  semesters  among  which  the  required  ten  hours  were  divides . Dur- 
ing this  time  I have  worked  with  carbohydrates  and  herbs  from  Peru, 
South  America.  In  the  first  case  levulinic  acid  and  furfural  were 
studied  and  in  the  latter  case  Peruvian  bark,  saponin  bark,  and  alga- 
rrobo  beans  were  studied.  The  last  part  of  my  work  which  constitutes 
the  isolation  of  the  alkaloid  or  alkaloids  in  algarrobo,  was  investi- 
gated becaui.se  of  the  physiological  properties  of  the  herb,  as  claimed 
by  the  natives  of  Peru  who  use  it  as  a stimulant. 

The  work  has  not  been  completed.  However,  there  have  been 
obtained  some  results  that  should  be  of  value  for  anybody  interested 
in  a further  work  with  the  same  material. 


Digitized  by  the  Internet  Archive 

in  2015 


https://archive.org/details/investigationofaOOmora 


1 


THEORETICAL  PART 
Levulinic  Acid 

The  yield  of  the  acid  was  studied  according  to  the  meth- 
ods of  W.A,  Noyes  in  his  "Manual  for  Organic  Chemistry",  and  Vanino's 
"Handbuch  Der  Praparativen  Chemie".  In  the  first  method  cane  sugar 
was  hydrolysed  by  hydrochloric  acid  and  water  into  fructose  and  glu- 
cose f ollowed  by  their  decomposition  into  levulinic  and  formic  acids 
and  water.  In  the  second  case  potao  starch  was  used  and  hydrolysed  by 
a dilute  solution  of  hydrochloric  acid.  Ihave  found  that  W.A. Noyes' 
method  gave  a better  yield  than  Vanino's. 

Furfural 

The  Eastman  Kodak  Co.  referred  to  the  university  that 
they  could  not  obtain  more  than  five  percent  of  furfural  from  corn 
cobs.  This  was  five  percent  less  than  the  percent  claimed  by  the  uni- 
versity. For  this  reason  three  checks  were  run  on  the  preparation  of 
furfural  according  to  the  directions  published  by  Dr.  R.  Adams,  Dr. 

0.  Karnm  and  Dr,  C.  S.  Marvel  in  a bulletin  entitled  "Organic  Chemical 
Reagents,  The  results  were  satisfactory  inasmuch  as  nine  and  three 
tenths  percent  was  obtained. 

Algarrobo 

This  bean  was  identified  by  Dr.  Trelease  of  the  depart- 
ment of  Botany  as  belonging  to  the  Prosonis  Horrida.  It  is  a yellow 
bean  about  six  to  seven  inches  long,  three  eighths  ofan  ich  wide  and 
one  fourth  of  an  inch  thick.  It  contains  a dark  brown  seed  similar 
to  those  of  apples,  and  has  a sweet  smell.  As  stated  in  the  intro- 
duction the  bean  was  studied  so  as  to  determine  the  substance  that 
had  the  stimulant  property.  It  was  supposed  that  the  bean  would  con- 


tain  an  alkaloid  of  very  good  physiological  properties.  With  these 
ideas  in  mind,  the  work  of  investigation  was  undertaken.  Although 
the  results  are  not  the  best,  an  approximation  to  the  proper  method 
of  extraction  has  been  accomplished.  Four  methods  of  extraction  were 
tried  before  anything  of  the  alkaloid  w as  seen. 

EXPERIMENTAL  PART 
Levulinic  Acid 

A liter  flask  was  used.  In  this  flask  one  hundred  cubic 
centimeters  of  water  and  one  hund.red  grams  cf  cane  sugar  were  mixed 
and  heated  on  the  steam  bath  for  twenty  hours,  with  a condenser  at- 
tached to  the  flask.  After  this  time  the  mixture  was  filtered  and  the 
residue  boiled  with  one  hundred  cubic  centimeters  of  water  and  fil- 
tered also.  To  the  combined  filtrates  thirty-five  grams  of  sodium 
hydroxide  were  added  in  solution.  This  alkaline  solution  was  evapo- 
rated to  about  one  hundred  cubic  centimeters,  filtered  and  extracted 
five  times  with  fifty  cubic  centimeter  portions  of  ether.  The  ether 
was  distilled  off.  The  residue  was  distilled  under  diminished  pre- 
ssure, and  the  fraction  boiling  at  140-L60  degrees  centigrade  and 
twenty-five  millimeters  pressure  was  collected.  The  distillate  was 
frozen  in  a wide  mouth  bottle.  The  watery  residue  was  separated  and 
the  res  warmed  to  room  temperature  and  weighed.  The  first  trial  gave 
twelve  grams  of  levulinic  acid,  and  the  second  one  seventeen. 

In  page  142  of  Vanino 's  second  volume,  the  following  di- 
rections were  found:  Mix  one  liter  of  hydrochloric  acid  sp,  gr.  1.1 

with  1 kilogram  of  starch  to  a syrup  and  then  reflux  the  mixture  for 
twenty  hours.  Cool,  filt  er,  and  distill  under  vacuum.  The  remainder 


3 

was  purified  in  the  same  way  a.s  the  preceeding  one.  By  this  method 
138  grams  of  acid  were  obtained.  The  previous  method  gives  a higher 
nercent  yield. 

Furfural 

As  already  stated  the  obiect  of  working  with  furfural 
was  to  check  the  yield  of  the  substance  from  corn  cobs.  The  process 
was  as  follows:  In  a 12  1 . round  bottom  flask  fitted  with  a cork 

stopper  holding  500  cc  separatory  funnel  and  a condenser  set  down- 
ward for  distillation,  are  placed  1300  grams  of  corn  cobs  (ground  to 
about  the  size  of  corn  kernels),  1800  cc  of  commercial  concentrated 
hydro chloric  acid,  and  46C0  cc  of  water.  After  standing  for  two  to 
three  hours,  the  flask  was  shaken,  and  heat  applied.  As  the  disti- 
llate comes  over,  more  acid  solution  is  added  ( 1 l.of  acid  to  15  1. 
of  water  )The  presence  of  furfural  is  shown  by  an  immediate  test  of 
of  a few  drops  of  the  distillate  on  a piece  of  paper  containing  some 
aniline  acetate.  This  teste  is  very  delicate.  The  distillat  e is  then 
treated  with  sodium  hydroxide  to  slight  acidi  ty,  and  then  salted 
end  distilled  until  the  distillate  does  not  give  a test  for  furfural 
with  aniline  acetate.  The  first  portions  coming  over  are  collected 
until  the  liquid  is  uniform. 

The  wet  furfural  is  now  distilled  under  diminished  pre- 
ssure using  an  oil  bath  the  temperature  of  which  should  not  be  allow- 
ed to  rise  above  130  degrees  centigrade.  The  first  distillate  consits 
of  water  and  furfural  but  after  about  60  cc  have  distilled,  purefur- 
fural  distils  over.  It  boils  at  90  degrees  centigrade  at '65  mm.  pre- 
ssure. The  yield  of  the  first  batch  was  9.3  percent,  and  of  the  other 
runs  8 percent.  The  last  two  runas  v;-ere  partly  spoiled. 


4 


Peruvian  Bark 

In  order  to  determine  the  percent  of  quinine  in  the  bark 
the  following  method  was  used:  The  bark  was  ground  to  almost  a 
powder  and  to  a poind  of  it  ninety  grams  of  lime  in  90C  cc  of  water 
were  added.  The  mixture  was  stirred  well  and  then  dried  in  a steam 
bath.  The  dry  mixture  was  then  extract  ed  with  800  cc  of  chloroform 
for  12  hours.  At  the  end  of  this  period  the  chloroform  was  filtered 
and  a fresh  amount  added,  600  cc.  The  combined  extracts  were  shaken 
with  dilute  sulphuric  acid,  225  cc.  After  separating  the  acid,  ano- 
ther portion  of  150  cc  was  added.  The  remaining  chloroform  solution 
was  washed  with  water  until  no  coloration  was  noticed.  The  combined 
water  and  acid  solutions  were  neutralized  bjr  ammonium  hydroxide,  and 
evaporated  until  crystals  began  to  form.  At  this  point  the  solution 
was  cooled,  and  the  crystals  separated.  The  remaining  solution  wa3 
once  more  evaporated  and  the  crystals  were  sepated.  In  this  way  3.85 
percent  of  quinine  was  obtained,  a sample  directly  imported  from 
Peru  was  run  through  the  same  process,  and  3.15  percent  of  cuinine 
was  obtained . 

A soxhlet  extraction  was  run,  which  did  not  come  out  very 
well.  It  seems  that  the  refluxing  to  which  the  chloroform  is  submittd 
a„ffects  the  quinine  somehow.  The  y ield  increased  by  five  tenths  per- 
cent, but  this  result  is  not  very  sure  because  the  melting  point  was 
not  satisfactory. 


Saponin  Bark 

The  following  was  the  method  of  extraction  for  saponin: 
Extract  three  times  with  water  the  ground  bark,  boiling  for  5 hours 
eanh  time.  The  combined  extract  was  evaporated  to  dryness.  The  resi- 
due was  ground  and  extracted  with  80  percent  alcohol.  The  alcohol 


- 


o 

extraction  was  continued  for  four  hours  according  to  Sttttz.  However, 
although  the  percent  of  the  crude  saponin  was  found  to  be  over  35 
percent,  the  directions  for  its  purification  did  not  yield  any  amount 
saponin.  Different  samples  were  tried  of  50  grains  and,  25  grams,  but 
in  each  case  no  results  were  obtained.  For  50  grams  of  extract  1000 
cc  of  80  percent  alcohol  were  used. 

In  the  first  extraction  the  liquid  was  cooled  before 
filtering,  and  a crystalline  prod„uct  was  obtained,  while  in  the  se- 
cond extraction  the  liquid  was  filtered  while  hot,  and  a gummy  ex- 
tract was  obtained,  which  was  hard  to  handle. 

While  the  bark  was  boiling  I noticed  a white  crystalline 
substance  sepatating  at  the  level  of  the  liquid  which  seems  to  be 
pure  sapbnin* 

Algarrobo 

After  reading  several  general  methods  of  analysis  for 
herbs  containing  alkaloids,  the  following  one  was  used..  The  algarro- 
bo bean  was  ground  as  fine  as  possible  ( 20  mesh  ) , then  it  was  ex- 
tracted with  ether  in  a soxhlet  ext  raetor  for  twenty  hours.  The 
ether  solution  had  a light  yellow  color  at  the  end  of  this  time.  Tien 
the  ether  solution  was  shaken  with  three  fifty  cc  portions  of  a 10 
percent  hydrochloric  acid  in  a separatory  funnel.  The  combined  por- 
tions of  acid  were  neutralized  with  ammonium  hydroxide,  and  this 
alkaline  solution  was  shaken  with  three  50  cc  portions  of  ether  and. 
evaporated  todryness.  There  was  a,  very  small  resid.ue,  which  was  taken 
up  by  one  cc  of  a five  percent  hydrochloric  acid.  The  acid  solution 
was  afterward  tested  with  Mayer's  reagent.  No  precipitate  was  seen. 
After  this  failure  to  obtain  any  alkaloid  with  an  ether 
extraction,  an  alcohol  extraction  was  tried.  The  ground  bark  was 


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6 


placed  in  a percolator  and  percolated  with  three  150  cc  portions  of 
hot  92  percent  alcohol.  The  combined  extract  solutions  were  filtered 
and  evaporated  to  a thick  syrup.  Then  the  syrup  was  treated  with  a 
five  percen  hydrochloric  acid,  and  left  to  stand  for  forty-eight 
hours  to  dewdsit  resinous  matter.  The  solution  was  filtered  after- 
wards and  because  it  was  too  dark  in  color,  it  was  clarified  with 
lead  acetate.  The  lead  was  precipitated  with  hydrogen  sulphide,  fil- 
tered, and  the  hyd.rogen  sulphide  precipitation  repeated  two  more 
times  , in  order  to  insure  a total  separation  of  the  lead.  To  the 
clear  solution  ammonium  hydroxide  was  added  in  excess.  Here  no  pre- 
cipitate was  obtained,  but  instead  a slight  change  in  color. 

The  alkaline  liquid  was  shaken  in  s separatory  funnel, 
with  four  50  cc  portions  of  ether,  50  cc  portions  seemed  to  do  an 
efficient  extraction.  The  ether  extract,  which  was  clear,  was  run 
into  a separatory  funnel  and  left  standing  to  separate  water.  It  was 
finally  dried  with  calcium  chloride  and  evaporated  under  diminished 
pressure.  The  residue  was  taken  up  by  1.5  cc  of  water  sligtly  acidi- 
fied with  hydrochloric  acid.  One  fifth  of  this  small  amount  was 
tested  with  Mayer's  reagent  which  gave  a very  good,  test  for  alka- 
loids. The  precipitate,  which  was  slightly  yellow,  turned  brownish 
on  standing.  This  was  a proof  of  the  presence  of  alkaloid  in  the  Al- 
garrobo  bean.  However,  its  solubility  in  alcohol  seemed  to  be  very 
little. 

A water  percolation  was  tried  next.  The  water  extract 
was  run  through  the  following  process:  It  was  made  alkaline  and  then 

extracted  in  a separatory  funnel  with  three  6 0 cc  portions  of  ether,. 
The  ether  was  extracted  with  dilute  h3rd.r0 chloric  acid.  The  a.cid 
solution  was  made  alkaline  with  ammonium  hydroxide  and  then  it  was 


7 


extracted,  with  ether.  The  ether  solution  which  was  sunnosed,  to  con- 
tain the  alkaloid,  was  evaporated  to  dryness.  The  residue  was  taken 
up  by  four  cc  of  acid  and  a fraction  tested  with  Mayer's  reagent. 

The  test  was  very  good,  and  it  revealed  that  the  alkaloidwas  more 
soluble  in  water  than  in  alcohol.  Theis  not  only  because  of  the 
amount  of  precipitate  but  also  by  the  amount  of  residue  left  after 
the  ether  was  wvaporated  to  dryness. 

Another  process  was  then  tried.  150  grams  of  ground  bean 
were  treated  with  a solution  of  10  grams  of  lime  and  one, hundred  cc 
of  water.  The  mixture  was  stirred  well,  dried  in  a steam  bath,  re- 
ground, and  extracted  with  ether  in  a soxhlet  extractor  for  twenty 
hours.  The  ether  extraction  was  treated  as  in  the  first  trial.  After 
the  ether  was  dried  and  distilled  under  diminished  pressure,  an  oily 
residue  was  left, about  0.7  cc,  and  about  three  white  crystals  of  the 
size  of  a sugar  granule  were  also  found.  It  was  questioned  whether 
this  oily  liquid  could  be  some  fat  extracted  from  the  bark.  The  a- 
mounts  were  so  small  that  no  constants  -could  be  investigated.  As  a 
conclusion,  it  can  be  said  that  there  is  alkaloid  in  the  bran,  but 
that  the  percent  is  very  small,  also  that  the  alkaloid  is  in  a form 
more  soluble  in  water  than  in  any  other  solvent,  and  that  only  the 
lime  treatment  renders  it  soluble  in  ether. 

In  as  much  as  the  bean  was  very  sweet  an  analysis  to  de- 
termine trie  kind  of  sugar  "'as  run.  An  osazone  of  the  unknown  sugar 
was  prepared  which  melted  at  the  melting  point  of  maltose.  For  this 
reason  the  osazone  of  pure  maltose  w as  prepared,  its  melting  point 
taken,  and  also  mixed  melting  points.  All  the  melting  points  werethe 
same.  This  indicated  that  maltose  was  the  sugar  in  the  bean.  Besides, 
a microscopic  examination,  where  the  crystals  of  the  osazone  of  the 


8 


bean  corresponded  to  pictures  of  the  crystals  of  pure  maltosazone. 

SUMMARY 

Summing  up  all  of  the  work,  it  has  been  found  that  ia  the 
case  of  furfural  the  distillate  from  the  flask  where  the  corn  cobs 
and  the  acid  solution  boils,  always  gives  a test  for  furfural  irres- 
pective of  the  time  of  distillation.  This  seems  to  indicate  that  the 
process  can  be  improved,  in  order  to  obtain  a larger  percent  of 
furfural , 

Peruvian  bark  is  an  herb  that  has  been  studied,  and,  in 
my  case,  I only  worked  on  the  percent  of  quinine  in  the  vark  dirctly 
imported  from  Peru.  In  spite  of  the  fact  that  the  soxhlet  ectraction 
of  the  bark  was  not  satisfactory,  I believe  that  a similar  process 
with  slight  qualifications  as  to  temperature  and  air  exposure  of  the 
apparatus,  so  as  not  to  have  to  heat  the  chloroform  so  much,  can  be 
divised.  If  the  chloroform  could  be  boiled  under  diminished  pressure 
while  subject  to  the  soxhlet  extraction,  this  would  prevent  any 
chemical  action  during  the  extraction,  for  this  would  render  a lower 
boiling  point. 

Regarding  saponin,  not  much  work  was  done  with  it  except 
to  follow  the  method  outlined  by  Siitz.  However,  there  is  a field  of 
study  in  the  purification  of  the  saponin* 

In  the  case  of  algarrobo,  Ihave  given  a detailed  explana- 
tion of  the  process  and  results  above.  I just  wish  here  to  restate 
the  fact  that  the  process  of  extraction  of  the  alkaloid  or  alkaloids 
can  be  improyed.  in  orderto  obtain  a*  larger  yield.  A further  study  of 
this  bark  would  reveal  important  things,  because  the  only  man  who 
worked  with  it  in  Peru,  although  he  did  not  carry  a chemical  analysis 
prepared  a bevef  age  which  he  labled  ''Algarrobina" , and  which  had  a 


r 


- 


. 


very  wood  tonic  effect.  This  medicine,  due  to  the  crude  way  in  which 
it  was  prepared,  and  due  as  st&ted,  to  the  lack  of  more  knowledge  of 
the  chemical  character  of  the  active  subs face , does  not  have  a large 
market*  It  was  my  intention  to  furnich  a complete  chemical  analysis 
of  the  bark  and  further  more,  to  stud^r  the  physiological  activity  of 
the  alkal&id.  However,  Ihave  not  accomplished  these  aims  entirely, 
but  in  part.  Because  the  percent  is  very  low,  it  can  be  inferred 
that  the  alkaloid  must  indeed  be  very  active. 

The  seeds  do  not  contain  any  alkaloid. 

BIBLIOGRAPHY 

Levulinic  acid: 

W.  A Noyes'  "Manual  for  Organic  Chemistry" 

Vanino's  "Handbuch  Der  Praparativen  Chemie" 

Furfural ; 

Bulletin  No.  43  of  the  University  of  Illinois. 

Peruvian  Bark  , Saponin  Bark,  and  Algarrobo  benns: 

"Chemistry  and  Analysis  of  Drugs  and  Medecines"  by 
Henry  C.  Fuller,  Pant  II  Chapter  III 

"Allen's  Commercial  Organic  Analysis"  Fourth  edition 
volume  VI  Pages  167  - 185. 

"Uber  Das  Saponin"  by  Dr.  Ed,  Stiitz  Pages  231  - 257. 


